Chemical modification of gamma-carboxyglutamic acid residues in prothrombin elicits a conformation similar to that of abnormal (des-gamma-carboxy)prothrombin.
Chemical modification of gamma-carboxyglutamic acid (Gla) residues in human prothrombin to gamma-methyleneglutamic acid (gamma-MGlu) residues elicited a conformation similar, if not identical, to that of des-gamma-carboxy prothrombin or PIVKA-II, i.e., prothrombin molecules induced by vitamin K antagonists or vitamin K deficiency states. The reaction seems to proceed sequentially by preferentially modifying a Gla at residue 32 that is located innermost among 10 Gla residues of human prothrombin. The initial modification resulted in nearly 50% losses of barium salt adsorption, the procoagulant activity and thrombin generation by the prothrombinase complex. The subsequent modification of two Gla residues at positions 6 and 16 gave rise to the immunoreactivity to an established monoclonal antibody that specifically recognizes the des-gamma-carboxy prothrombin. Further modification of Gla residues increased the reactivity to the antibody, indicating that the conformation recognized by the antibody was stabilized so as to more readily fit the recognition site of the antibody. The appearance of the immunoreactivity was obviously related to the modification of Gla residues in prothrombin, since all other similarly treated derivatives of prothrombin lacking the Gla-domain failed to react with the antibody. Such chemically modified prothrombins may serve as models for studying abnormal des-gamma-carboxy prothrombin produced in vitamin K deficiency states.
Sugo
T
T
Institute of Hematology, Jichi Medical School, Tochigi.
Watanabe
K
K
Naraki
T
T
Matsuda
M
M
D015055:1-Carboxyglutamic Acid / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D011498:Protein Precursors; D011516:Prothrombin / Q000031:analogs & derivatives / Q000737:chemistry* / Q000276:immunology; D014813:Vitamin K Deficiency
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D011498:Protein Precursors; C008208:acarboxyprothrombin; D015055:1-Carboxyglutamic Acid; D011516:Prothrombin